Conjugated unsaturated glyceride mixtures and a method for producing the same

ABSTRACT

The invention relates to a process for conjugating two double bonds separated by a methylene bridge in a long-chain alkene, in which the long-chain alkene is isomerized with an
         a) imino base, or   b) iminophosphorane base,
 
as catalyst.

The present invention relates to processes for preparing conjugatedalkenes, in particular esterified fatty acids. The inventive process canpreferably be used to prepare glycerides of conjugated fatty acids fromsynthetic or natural glyceride oils or fats or derivatives thereof. Thepresent invention further relates to glyceride mixtures obtainable bythe inventive process and to preparations which comprise the inventiveglyceride mixtures. The invention further relates to processes forpreparing the inventive preparations, processes for preparing drugs andkits which comprise the inventive preparations and/or glyceridemixtures.

TECHNICAL FIELD

Fatty acids and triglycerides have a multiplicity of applications in thefood industry, animal nutrition, cosmetics and in the pharmaceuticalsector. Depending on whether these are free saturated or unsaturatedfatty acids, or triglycerides having an elevated content of saturated orunsaturated fatty acids, they are suitable for the most widespreadapplications. Thus, for example, a high content of lipids havingunsaturated fatty acids and, especially, polyunsaturated fatty acids, isimportant for the nutrition of animals and humans, since these, forexample, have a beneficial effect on the triglyceride level orcholesterol level and thus decrease the risk of a heart disorder.Unsaturated fatty acids are used in various dietetic foods or drugs.

Particularly valuable and sought-after unsaturated fatty acids are theconjugated unsaturated fatty acids, for example conjugated linoleic acid(CLA). Conjugated polyunsaturated fatty acids are somewhat rare comparedwith other polyunsaturated fatty acids.

CLA is a collective term for positional and structural isomers oflinoleic acid that are characterized by a conjugated double bond systemstarting at carbon 7, 8, 9, 10 or 11. Geometric isomers exist for eachof these positional isomers, that is cis-cis, trans-cis, cis-trans,trans-trans.

C18:2 cis-9, trans-11 and C18:2 trans-10, cis-12 CLAs which are theisomers with the highest biological activity are especially ofparticular interest, since in animal experiments they have proved to becancer-preventive, to have antiarteriosclerotic activity and, in humansand animals, to reduce the body fat content. Commercially, CLAs arecurrently principally marketed as the free fatty acid.

For humans, the most important natural sources of CLA are especiallyanimal fats. Thus fats of ruminant animals such as cattle (Chin, Journalof Food Composition and Analysis, 5, 1992: 185-197) and sheep, and alsodairy products, have very high CLA concentrations. In cattle from 2.9 to8.9 mg of CLA/g of fat are found. In contrast, vegetable oils,margarines and fats from nonruminant animals have CLA concentrations offrom only 0.6 to 0.9 mg/g of fat.

BACKGROUND ART

A number of beneficial-effects have been detected for CLA. Thus, theadministration of conjugated linoleic acid decreases the body fat inhumans and animals and improves the feed utilization in animals (WO94/16690, WO 96/06605, WO 97/46230, WO 97/46118). By administeringconjugated linoleic acid, for example, allergies (WO 97/32008), diabetes(WO 99/29317) or cancer (Banni, Carcinogenesis, Vol. 20, 1999:1019-1024, Thompson, Cancer, Res., Vol. 57, 1997: 5067-5072) may also bebeneficially affected. Polyunsaturated fatty acids are also added toinfant food to “increase the nutritional value” and as essentialbuilding blocks which ensure growth and brain development.

Since CLA only occurs naturally in significant quantities in ruminantsand their products, such as milk, cheese, etc., there is a great needfor alternatives to CLA originating from these animal sources, in orderto ensure balanced and healthy nutrition, in particular if the supplywith animal fats is reduced, inadequate or if synthetic preparation istoo expensive. Commercially, CLAs are currently principally marketed asfree fatty acid. Free fatty acids such as CLA, however, generally do notoccur naturally as free fatty acids, but are esterified to formbiologically active triglycerides. In addition free fatty acidsfrequently possess disadvantageous sensory properties. For incorporationinto foods, for example, triglycerides are also preferred to free fattyacids for technological reasons.

The processes described in the prior art generally consist of two orthree process steps in which free fatty acids or their alkyl esters arefirst prepared and isomerized in order then to transesterify these withglycerol or glycerides under enzymatic or chemical catalysis to formtriglycerides.

In the conventional preparation method for free CLA acids, unconjugatedlinoleic-acid-containing oils (for example sunflower seed oil, soybeanoil or safflower oil) are isomerized, for example, with NaOH or KOH inethylene glycol at 180° C. (Ip C. et al., Cancer Res. 51 (1991)6118-6124). This process requires superstoichiometric amounts of alkali(based on fatty acids present in the oil) and produces substantialamounts of unwanted CLA isomers (in particular 8t, 10c- and 11c,13t-CLA).

EP-839897 describes a process in which, for the isomerization, linoleicacid-containing oils are reacted with KOH in propylene glycol at 150° C.Free CLA acids are obtained which contain only relatively small amountsof unwanted isomers. This process requires superstoichiometric amountsof KOH and corresponding amounts of mineral acids.

In one process, linoleic acid alkyl esters-are isomerized with catalyticamounts (0.3 to 1%) of potassium alkoxide, with CLA alkyl esters beingobtained (DE-1156788 and DE-1156789).

To prepare triglycerides, in EP-0779033, linoleic acid is isomerized at180° C. with NaOH in ethylene glycol to form free CLA and the free CLAis transesterified with palm oil triglycerides using immobilized Mucormiehei lipase. The triglyceride obtained as product containsapproximately 8% of each of the two wanted CLA isomers (9c, 11t- and10t, 12c-) in esterified form.

In some non-enzymatic processes, the esterification of glycerol and thetransesterification of natural fats and oils with free CLA acids iscarried out with the addition of known esterification catalysts at hightemperatures (from 180 to 240° C.) (Mikusch; Farben, Lacke,Anstrichstoffe 4 (1950) 149-159; DE-19718245). The resultantCLA-containing triglycerides, owing to the temperature stress necessaryin the process, have a high content of isomers which are unwanted fornutritional uses (in particular 8t,10c- and 11c,13t-CLA fatty acidradicals).

WO 01/18161 describes a solvent-free synthetic process for preparingCLA. The preparation of CLA from oil by alkali isomerization is alsodescribed there.

The individual CLA isomers can also be transesterified with the palm oiltriglycerides after enrichment of the isomers. In this manner a CLAcontent of 30% in the triglyceride can be achieved. (McNeill, J. Am. OilChem. Soc. 76 (1999) 1265). The transesterification of butter fat withfree CLA is based on a similar process, inter alia, immobilized Candidaantarctica lipase acting as preferred catalyst (Garcia, Biotechnol.Tech. 12 (1999) 369-373; Garcia, J Dairy Sci 2000, 83:371-377; Garcia,Biotechnology Letters (1998) 20:393-395). In a similar manner, corn oilwas also modified using chemically synthesized CLAs using lipasecatalysis (Martinez, Food Biotechnology 1999, 13:183-193).

Processes described in the prior art which start from CLAs in the formof free fatty acids for preparing the triglycerides thus, usingstoichiometric amounts of bases, release the fatty acids from oilscontaining non-conjugated polyunsaturated fatty acids and simultaneouslycarry out the conjugation. In the second process step, conjugatedpolyunsaturated fatty acids are reacted with glycerol or glycerides toform glycerides containing conjugated, polyunsaturated fatty acids.

Furthermore, in the prior art, in the preparation of glyceridescontaining conjugated, polyunsaturated fatty acids from glycerides orglycerol by reaction with alkyl esters of conjugated polyunsaturatedfatty acids, the alkyl esters of conjugated, polyunsaturated fatty acidsare obtained by transesterifying non-conjugated, polyunsaturated fattyacid-containing oils using catalytic amounts of alkali metal bases andsuperstoichiometric amounts of alcohols to give the alkyl esters ofpolyunsaturated fatty acids and these are then converted into the alkylesters of conjugated fatty acids in a second process step usingcatalytic amounts of alkaline earth metal bases.

The processes described in the prior art thus have the disadvantage thattwo to three process steps are required, which is an economicdisadvantage, to prepare glycerides containing conjugated,polyunsaturated fatty acids.

DISCLOSURE OF INVENTION

It is an object of the present invention thus to provide a rapid andeconomically advantageous process for producing conjugated double bondsin alkenes, in particular in esterified fatty acids, such as are presentin triglycerides.

We have found that this object is achieved by the embodiments underlyingthe present invention.

The present invention therefore relates to a process for preparing acompound of the formula (III)

from a compound of the formula II

which comprises reacting the compound of the formula IV with an

-   -   a) imino base, or    -   b) iminophosphorane base;    -   where n=1, 2, 3; and where        R¹ and R², or R¹ or R², can be:    -   H;    -   unbranched or branched C₁-C₁₀-alkyl,        -   where 0, 1, 2 or 3 carbons can be replaced by epoxide, O, S,        -   NZ, and/or —X¹—(C═X²)—,        -   where X¹ is a bond, O, S or NZ,        -   and/or X² is O, S or NZ;    -   mono-, bi-, or tricyclic aromatic saturated or partially        unsaturated C₀-C₆-alkyl carbocycle or heterocycle having from 3        to 6 carbons,        -   where from 0 to 3 heteroatoms can be selected from the group            consisting of S, N, and/or O;    -   and where each carbon of the alkyl chains or of the ring can        bear 1, 2 or 3 of the following substituents OZ, SZ, (C═O )—OZ,        NZZ¹, C₁ to C₆-alkyl, F, Br, Cl, I;    -   and where Z and/or Z¹ can be H or C₁-C₆-alkyl.

Particularly preferably, R¹ or R² independently of one another is H oran unbranched or branched C₁-C₁₀-alkyl, where each carbon of the alkylchains or of the ring can bear up to three of the following substituentsOZ, SZ, (C═O)—OZ, NZZ¹, C₁ to C₆-alkyl, F, Br, Cl, I, and where Z and/orZ¹ can be H or C₁-C₆-alkyl.

Preferably, the compound IV is a substituted or unsubstituted unbranchedalkene having from 8 to 24 carbons as described below.

More preferably, the alkyl chain bears one of the following substituentsOZ, SZ, (C═O)—OZ and Z and Z¹ as above, particularly preferably in aterminal position. Preferably, compound IV is a C8 to C24 fatty acid,with esterified fatty acids being particularly preferred.

The invention consequently also relates to a process for conjugating twodouble bonds separated by a methylene bridge in a long-chain alkene, inwhich the long-chain alkene is isomerized with an

-   -   a) imino base, or    -   b) iminophosphorane base,        as catalyst.

A “long-chain” alkene according to the invention is an unbranchedunsaturated carbon chain of at least six carbons. Preferably theinvention relates to alkenes which contain from 8 to 24, more preferablyfrom 10 to 24, preferentially from 12 to 22, even more preferably from16 to 20 carbons. The double bonds of the long-chain alkene areseparated from one another by a methylene bond (homoconjugated). Thealkene can also contain more than one double bond, provided that itcomprises one homoconjugated double bond. The term “alkene” alsocomprises derivatives of the alkene. Thus the alkene can be bound toother groups. Examples of such groups are described below for compound Ias R. These can be bound terminally to the alkene, for example.

An “imino base” is a base that has an═NR— group of atoms (imino group)and preferably an additional proton donor, for example the —PH—, orpreferably —NH—, group of atoms, in particular iminoamino bases. A“phosphorane” is an organophosphorus compound that has a valency of 5.An “iminophosphorane” additionally has an imino group, an“aminoiminophosphorane” additionally has up to three amino groups andone imino group and is also included. In particulartriamino(imino)phosphoranes are included.

In the prior art, the isomerization of non-conjugated, polyunsaturatedfree fatty acids or their alkyl esters is described using basiccompounds, such as alkoxides, alkalis, sodium amide or amines.

Houben Weyl, volume 5, pp. 63 ff. DE 25 17 377 describes theisomerization of 2,5-heptadienoic esters to their 3,5 derivatives usingstrongly basic amines, for example primary, secondary or tertiaryamines, or quaternary ammonium bases, at temperatures from 20° C. to 90°C. Zakharkin, 1982, describes the isomerization of the methyl ester of2Z,5E-heptadienoic acid to give the methyl ester of 3E,5E-heptadienoicacid, and of the 2E,5Z- and 2Z,5Z-ester mixture to give the methyl esterof 3E,5Z-heptadienoic acid (Zh. Org. Khim. 18 (1982) 11, 2261-66); inthis process catalytic amounts of triethylamine are used to isomerizethe double bond which is conjugated directly to the methyl ester. Theuse of the imino base guanidine and aminophosphorane bases totransesterify fatty acids in vegetable oils using protic alkyl alcohols,in particular methanol, to give their fatty alkyl esters is described bySchuchardt (J. Braz. Chem. Soc. Vol. 9, No. 3, 199-210, 1998;BR8202429).

The present invention is based on the surprising finding thatpolyunsaturated, non-conjugated double bonds, in particular inesterified fatty acids, can be isomerized to form conjugated doublebonds using catalytic amounts of certain very strong bases, that is tosay imino bases or iminophosphorane bases, in particular aminoiminobases or aminoiminophosphorane bases. The use of amines, in particularalso triamines, as has been described in the prior art, did not lead toan isomerization of homoconjugated double bonds to form conjugateddouble bonds in the esters of fatty acids studied according to theinventive process. The examples tabulate by way of example which amineswere used as catalysts for the inventive process.

Possibly, in the prior art, the double bond isomerized by triethylamineis activated by the adjacent carbonyl group. Preferably, therefore, noneof the double bonds to be isomerized is activated. The term“non-activated double bond” here is taken to mean that the double bondsof the long-chain alkene to be conjugated are separated from anactivating group by at least one methylene bridge. An activating groupis taken to mean a group of atoms that facilitates substitution of theadjacent double bond compared with a non-adjacent double bond. Thelong-chain alkyl can bear no activating groups, or one or moreactivating groups; preference is given to a terminal activating group,the activating groups being localized in such a manner that the doublebond to be isomerized is not activated. An activating group inparticular has a high electronegativity. Activating groups can be, forexample: —COOH, —COO—, —OH, —SO₄, —SO₃—, —CN—, ═NR—, or halogen (F, Br,Cl, I). Most preferably, the activating group is an alkyl ester.Preferred alkyl esters are described below. In a preferred embodimentcompound IV is therefore a fatty acid ester as described below.

The present invention therefore relates to a process for preparing aconjugated unsaturated fatty acid ester (FAE I), wherein apolyunsaturated fatty acid ester (FAE II) is isomerized using an iminobase or iminophosphorane base as catalyst, in particular using anaminoimino base or aminoiminophosphorane base. The inventive process canbe used, in particular, to convert alkyl esters of non-conjugatedpolyunsaturated fatty acids into their conjugated form.

A “fatty acid” is preferably an unbranched carboxylic acid having aneven number of carbons (n) (e.g. in n=16). Preferably the inventionrelates to fatty acids having from 8 to 24, more preferably from 12 to22, even more preferably from 16 to 22 carbons, particularly preferablyfrom 18 to 22 carbons, very particularly preferably having 18 carbons.

A “polyunsaturated fatty acid” is a fatty acid having at least twodouble bonds, which can be conjugated or non-conjugated. If notspecified otherwise, “polyunsaturated fatty acid” generally refers tofatty acids containing non-conjugated double bonds. A “conjugated,unsaturated fatty acid” is an unsaturated fatty acid having at least twodouble bonds that are conjugated. The non-conjugated, polyunsaturatedfatty acid has two double bonds which are at positions n and n+3 (thatis to say are homoconjugated), for example in the case of linoleic acidor linolenic acid, where n is a carbon of the carboxylic acid (seeabove).

MODE(S) FOR CARRYING OUT THE INVENTION

In an embodiment of the inventive process, the compound IV is present asesterified fatty acid (FA II), in particular as an alkyl ester.

An “alkyl ester” of the fatty acids is an ester thereof with alkanols,preferably with C₁- to C₅-alkanols, for example methanol, ethanol,propanol, isopropanol, n-butanol, isobutanol tert-butanol, or n-pentanoland its isomers (2-pentanol, 3-pentanol, 2-hydroxy-3-methylbutane).Particular preference is given to methanol and ethanol.

An “alkyl ester” of a fatty acid is also an organic compound so thatsaid alkanols can be bound to other carbons or heteroatoms, for exampleH, O, S, P, halogens. Also included are bonds to aromatic and cyclicorganic compounds and the derivatives enumerated below. Preference isgiven to alkyl esters which can be isomerized to form conjugatedpolyunsaturated fatty acids, for example to form conjugated linoleicacids (CLAs), α-parinaric acid (18:4 octadecatetraenoic acid),eleostearic acid (18:3 octadecatrienoic acid), dimorphecolic acid,conjugated linolenic acids and calendic acid, with particular preferencegiven to CLA preparations which comprise 9cis,11trans-CLA alkyl estersand 10trans,12cis-CLA alkyl esters as product.

In a preferred embodiment of the inventive process, the compound IV orthe esterified fatty acid (FA II) is present as a glyceride.

Surprisingly, it has been found that by means of the inventive processisomerization of polyunsaturated fatty acids, for example linoleic acid,that are esterified with a glyceride, in particular including with atriglyceride, it is possible to give conjugated unsaturated fatty acidscontained in a glyceride, without releasing the fatty acids. Asingle-stage isomerization of polyunsaturated free fatty acids which arepresent bound in a glyceride with catalysis by imino bases oriminophosphorane bases, in particular aminoimino bases oraminoiminophosphorane bases is not described in the prior art. To dateno economic process has been known for preparing glycerides containingconjugated polyunsaturated fatty acids in one stage with quantitativeconversion from glycerides containing non-conjugated, polyunsaturatedfatty acids. The inventive process, in contrast to the prior art, canstart directly from the oils or fats containing non-conjugated,polyunsaturated fatty acids and avoids chemical derivatization to givethe free fatty acids or their alkyl esters. The inventive process thusmakes possible direct conjugation of non-conjugated polyunsaturatedfatty acids directly in the glyceride.

A “glyceride” is glycerol esterified with one, two or three carboxylicacid radicals (mono-, di- or triglyceride). “Glyceride” is also taken tomean a mixture of different glycerides. Glyceride or the glyceridemixture can comprise other additives, for example free fatty acids,antioxidants, proteins, carbohydrates, vitamins and other substances, asare enumerated, for example, below under “additives”.

The glyceride used in the inventive process can also be present in asynthetic or naturally occurring glyceride oil or a derivative ormixtures thereof. “Glyceride” can also be taken to mean, depending onthe context, synthetic or naturally occurring fatty acid esters and/oroils and fats comprising glycerides, also referred to as “glyceridemixture” below.

“Glyceride” within the meaning of the inventive process is further takento mean derivatives derived from glycerol. In addition to the abovedescribed glycerides of fatty acids, these also includeglycerophospholipids and glyceroglycolipids. Preference is given here tothe glycerophospholipids such as lecithin (phosphatidylcholine),cardiolipin, phosphatidylglycerol, phosphatidylserine andalkylacylglycerophospholipids, such as plasmalogen. In particular,derivatives in which the fatty acid composition of thenaturally-occurring non-conjugated or saturated glycerides has notsubstantially changed are included.

Preferred starting materials are particularly glycerides or mixtures ofglycerides, in particular of mono-, di- or triglycerides, that areesterified with at least one, preferably two or three, polyunsaturated,in particular homoconjugated, fatty acids. Therefore, preference isgiven to synthetic or natural glycerides or glyceride mixtures whichcontain acyl radicals having from 1 to 22 carbons, preferably having 18carbons. Particular preference is given to natural oils and fats whichcontain polyunsaturated homoconjugated acyl radicals having more than 16carbons and less than 22 carbons, preferably from 18 to 20 carbons.

The term “oil” or “fat” is taken to mean a mixture of fatty acids thatcomprises unsaturated, non-conjugated, preferably homoconjugated,esterified fatty-acid(s), in particular linoleic acid. Preferably, theoil or fat has a high content of unsaturated, non-conjugated esterifiedfatty acid(s), in particular linoleic acid. Preferably, the content ofunsaturated, non-conjugated esterified fatty acids is approximately 30%,more preference is given to a content of 50%, still more preference to acontent of 60%, 70%, 80%, 90% or more. For determination, the fatty acidcontent can, for example, be determined by gas chromatography afterconverting the fatty acids into the methyl esters bytransesterification. The oil or fat can comprise various other saturatedor unsaturated fatty acids, for example calendic acid, palmitic acid,stearic acid, oleic acid etc. In particular, depending on thepreparation method, the content of the various fatty acids in the oil orfat can vary. Each fatty acid profile is included by the inventivepreparation, in particular fatty acid profiles which are produced in theproduction of oil from vegetable material. Preferably, the fatty acidesters are present as a glyceride, in particular as a triglyceride.

Preference is therefore given to an inventive process where theglyceride mixture can be of animal, microbial or vegetable origin, forexample olive oil, canola oil, coconut oil, coconut fat, sesame seedoil, rice germ oil, bamboo oil, bamboo fat, sunflower seed oil, rapeseedoil, fish oil, tallow oil, soybean oil, palm oil, safflower oil, linseedoil, wheatgerm oil, peanut oil, cottonseed oil, corn oil, pig fat, beeffat, poultry fat, milk fat, tung oil or shea oil or a derivative or amixture thereof. Particular preference is given in particular to oilsand fats which have a high content of linoleic acid, for examplesunflower seed oil, soybean oil, cottonseed oil, corn oil or wheatgermoil, safflower oil, thistle oil, rapeseed oil and in particular oils orfats from modified plant cultivars, in particular what are termed highlinoleic seeds, for example linola (from linseed oil). The modifiedplant cultivars can be bred or advantageously can also be produced bymutagenesis (for example GMO) (Angew. Chem. 2000, 112, 2292-2310).

The starting material for the inventive process can also be produced byconventional processes known to those skilled in the art, for exampleoil from plants. Oil can be produced by pressing, for example, seedhaving a high husk content, or husked seed. For pressing and production,in addition to vegetable seed, other plants parts, for example leaves,tubors, stems, blossoms, fruits etc. of suitable plants can also be usedwhich have a high content of unsaturated fatty acids, preferablyesterified in triglycerides. Whole plants can also be used. The pressedmaterial can also be pressed repeatedly.

Other materials which are also suitable for producing oils and fatssuitable for the inventive process are microorganisms, such asThraustochytrium or Schizochytrium strains, algae such as Phaeodactylumtricornutum or Crypthecodinium species, ciliates, such as Stylonychia orColpidium, fungi such as Mortierella, e.g. Mortierella alpina,Entomorphthora or Mucor. By means of strain selection, a number ofmutant strains of the corresponding microorganisms have been developedthat produce a series of desirable compounds, including PUFAs and whichare also suitable for producing said fatty acids or oils. In particularmicroorganisms can be produced by suitable transformations, for exampleusing nucleic acid molecules coding for desaturases or elongases.

Preferably, in the inventive process, linoleic ester is converted toconjugated linoleic ester (CLA).

Preference is therefore given to starting products which compriselinoleic esters, for example those which have a high content of linoleicacid-containing triglycerides. Particular preference is therefore givento processes in which natural oils and fats that have a high content oflinoleic acid, as described above, are converted according to theinvention, for example sunflower seed oil, soybean oil, safflower oil,linseed oil or derivatives of the same.

In an embodiment, in the inventive process, the catalysis is carried outusing the compound I

where independently of one another

-   -   X₁ can be —NH— or —PH—, preferably —NH—,    -   X₂ can be C, N or P, preferably N or P, most preferably N, and        where    -   R1 to R4 independently of one another can be:    -   H,        -   branched or unbranched C₁- to C₂₀-alkyl, where from 0 to 3            carbons can be replaced by O, S, NZ and/or —X₃—(C═X₄)—,        -   mono-, bi-, or tricyclic, aromatic, saturated or partially            unsaturated C₀- to C₆-alkylcarbocycle or heterocycle having            from 3 to 17 carbons, where from 0 to 3 heteroatoms can be            selected from O, S, NZ and/or —X₃—(C═X₄)—;        -   and where each carbon of the alkyl chains or of the ring can            bear up to three of the following substituents OZ, SZ,            (C═0)—OZ, NZZ₁, C₁- to C₆-alkyl;    -   where X₃ can be a bond, O, S or NZ, and/or X₄ can be O, S or NZ;        and    -   where Z and/or Z₁ independently of one another can be H or C₁-        to C₆-alkyl.

X1 and X2 can thus be part of a ring via R3 or R4, in particular R1 andR4, and R2 and R3, can be part of a ring. The rings can therefore bearheteroatoms or further double bonds. In particular, R1 and R4, and R2and R3, can be cyclically linked via (CH)₂n, where n=2, 3, 4, 5.

It is known to those skilled in the art that the use of compound I whereX₁ is —PH— is only possible under inert conditions that preventoxidation of —PH—.

Compound I can also be polymer-bound, for example as Merrifield resins,for example heterogenized as Merrifield resins on variouschloromethylated poly(styrene/divinylbenzene) resins, or heterogenizedon polystyrene resin after introduction of a spacer in the form of analkyl chain.

The catalyst can be present both as pure substance and immobilized, forexample bound to a polymer (polymer-bound guanidine bases (J. Mol.Catal. A: Chemical 109 (1996) 37-44; Pure Appl. Chem., A29(3), 249-261(1992)), polymer-bound aminoiminophosphorane bases (Chimia 39 (1985)No.9, 269-272) or incorporated into a support (cyclohexylguanidine inzeolite Y; THL, Vol. 38, No. 8, 1325-1328, 1997)).

Preferably, compound I has the following structure (Ia):

where X is C, N, or P, preferably N or P, most preferably N, and R is asdefined above for R1 to R4.

Preference as catalyst is also given to compounds (II):

More preference is given to compounds (IIa):

More preference is given to compound (IIb):

where in compounds (II) to (IIb), for R and R1 to R6, the same appliesas above for R1 to R4.

In particular, R1 and R2, R3 and R4, and/or R5 and R6 can be cyclicallylinked.

Surprisingly, it was found in the inventive process that compound I orII, in particular imino bases or iminophosphazene bases, preferablyaminoimino bases or aminoiminophosphazene bases, can catalyze theisomerization of non-conjugated polyunsaturated fatty acids in theglyceride without addition of protic solvents, for example alkylalcohols. Particular preference is therefore given to carrying out theinventive process using guanidine bases, or the bases enumerated below,as catalyst.

Particular preference is given to 1,5,7-triazabicyclo[4.4.0]dec-5-ene(TBD) or analogous diaza bases, for example1,2,3,4,4a,5,6,7-octahydro-1,8-naphthyridine [CAS 60832-40-8]. Mostpreference is given to TBD.

Particular preference is also given to the phosphazene base P4-T-BU [CAS111324-04-0], phosphazene base P1-T-Oct No. [CAS 161118-69-0],phosphazene base P1-T-Bu-tris(tetramethylene) [CAS 161118-67-8],phosphazene base P2-T-Bu [CAS 111324-03-9], phosphazene base P4-T-Oct[CAS 153136-05-1], the salts1,1,1,3,3,3-hexakis(dimethylamino)diphosphazenium fluoride [CAS137334-99-7], 1,1,1,3,3,3-hexakis(dimethylamino)diphosphazeniumtetrafluoroborate [CAS 137334-98-6] or2-tert-butylimino-2-diethylamino-1,3-dimethylperhydro-1,3,2-diazaphosphorine[CAS 98015-45-3].

The above salts (for example BF₃ ⁻, or F⁻ salt) or other salts of saidcompounds are also claimed.

Preferred solvents for carrying out the inventive process are nonprotic,for example ethyl esters, hexane, heptane, DMSO, DMF, MTBE, or THF.However, those skilled in the art, on the basis of the disclosure of thepresent invention, can find by simple test series other solvents whichcan also advantageously be used in the inventive process. The inventiveprocess, however, can also be carried out with the addition of proticsolvents, for example alkyl alcohols.

Conjugated fatty acids and their esters are preferably used in drugs orfoodstuffs for humans and animals and in cosmetics. Therefore, it isadvantageous to carry out the inventive process using solvents which arenontoxic, or are of as low toxicity as possible, so that it is notnecessary to separate off the solvent after carrying out the process, orslight traces are not hazardous for the desired use.

Depending on use, a different solvent can be advantageous. For exampleprotic solvents, for example methanol, ethanol etc., can also be used,for example for conjugating esters of unsaturated homoconjugated fattyacids. It can be advantageous to use a solvent which can be readilyseparated off from the fatty acids, glycerides, oils or fats, forexample can be extracted by shaking in water or which is low-boiling,for example MTBE.

Advantageously, the inventive process can be carried out withoutsolvent. The reaction temperature of the process must then be adapted sothat the melting point of the catalyst used is exceeded. It can beadvantageous to carry out the process at low temperatures and todissolve the catalyst in a suitable solvent for this. However, fromeconomic aspects, it can also be advantageous not to use solvent.

Advantageously, the inventive process is carried out at temperaturesbelow 180° C. High temperatures in the isomerization lead to loweryields, for example owing to derivatizations or destruction of the fattyacids. The temperature of the inventive process can be chosendifferently, depending on solvent, catalyst, pressure and startingmaterial.

The temperature and reaction time depend, for example, on the strengthof the base used. Thus TBD is a strong guanidine base, but the reactiontemperature, owing to the high melting point, must be chosen to be above130° C. if no solvent is used. Preferably the temperature in the caseof, for example, guanidine bases such as TBD without solvent, is abovethe melting point but below 200° C., for example from 120° C. to 180°C., more preferably from 120° C. to 160° C., still more preferably from130° C. to 140° C. The reaction times should be chosen accordingly. If asolvent is used the reaction temperature and reaction time can beadapted. Via simple series of experiments those skilled in the art canestablish the yield and the amount of by-products, in particular ofunwanted isomers, and harmonize the experimental conditions accordingly.The same applies to employing other catalysts used according to theinvention. Thus the phosphazene bases, depending on conditions, react atas low as 0° C., but, from energetic aspects, higher temperatures arepreferred, for example room temperature. Those skilled in the art couldaccordingly exchange the base selected, change the temperature or adaptthe reaction time, if too many by-products occur. It is thought that theamount of by-products, in particular the trans/transfatty acids, iscaused by a frequent change between deprotonation and protonation. Theexamples show how those skilled in the art can derive suitableexperimental conditions from the combination and variation of variousparameters.

Furthermore, the inventive process can be carried out continuously orbatchwise.

The inventive process can be carried out not only batchwise, but alsocontinuously. For the continuous procedure, a tubular reactor, forexample, can be used, in which the catalyst capable of isomerization ispresent. The catalyst can be present not only as pure substance, butalso immobilized, for example bound to a polymer (polymer-boundguanidine bases (J. Mol. Catal. A: Chemical 109 (1996) 37-44; Pure Appl.Chem., A29(3), 249-261 (1992); polymer-bound aminoiminophosphorane bases(Chimia 39 (1985) No. 9, 269-272) or incorporated into a support(cyclohexylguanidine in zeolite Y; THL, Vol. 38, No. 8, 1325-1328,1997)). Continuous reaction of vegetable oils with polymer-boundguanidine bases is described in BR 8202429.

In addition, a process is included according to the invention in whichthe catalyst is separated off in a further step.

In order that the product of the inventive process can be used as a foodadditive for humans and animals, it can be expedient to separate off thesolvents and catalysts used.

The solvent can be separated off by distillation or removal by aqueousextraction. The same applies to the catalysts.

In one step, in the inventive process, the fatty acid can be cleavedoff. Also, in one step of the process, the fatty acid can betransesterified. Corresponding processes are known to those skilled inthe art and are described above. These steps can be carried outsubsequently or simultaneously with the described process steps.

In a further embodiment, the present invention relates to the product ofthe inventive process, in particular to a conjugated alkene, for examplein particular in the form of a glyceride, but also in other forms, asare included under the above description for “alkenes”, for example asfatty acids, esterified fatty acids, for example glycerides, alsomixtures of different alkenes which are used as starting material, or aglyceride mixture obtainable by the inventive process, the product stillbeing able to contain e.g. the catalyst, preferably in nontoxic amounts.

The inventive product comprises the abovementioned catalyst, for exampleeven only in traces, or has been prepared from a natural glyceridemixture, for example one of the abovementioned oils or fats, or amixture or a derivative thereof, or from a complex synthetic oil or fat.

“Complex synthetic oil or fat” is taken to mean those oils or fats whichconsist of more than one glyceride or of one or more mono- ordiglycerides, for example of glycerides that have at least one identicalor different fatty acids bound in one or different esters of theglyceride, or that have at least two different fatty acids, or the fattyacids of which are bound to different carbons of the glyceride (forexample 1,2 and 1,3 glycerides), or triglycerides of at least twodifferent fatty acids.

Complex synthetic oils or fats are, in particular, those oils and fatsthat have glycerides or mixtures of glycerides which, after cleavage ofthe fatty acids, can no longer be prepared in the same structure and inthe same composition. The processes described in the prior art makepossible the synthesis (isomerization) of polyunsaturated fatty acidsthat are esterified with a glyceride only after cleavage ortransesterification, conjugation and reesterification with glycerides orglycerol. As a result the structure of the glycerides in the preparedmixtures, oils or fats and their composition change. Glyceride mixtureswhich are obtainable by the inventive process, however, have essentiallythe same composition as the starting mixture, but the polyunsaturatedfatty acids (for example linoleic acid), owing to the inventive process,are conjugated to an increased extent (for example to form CLA).Advantageously, in particular, the CLA content in the product isincreased. The composition of the residual fatty acids and, inparticular, of the non-conjugated fatty acids esterified withglycerides, is essentially unchanged in the product.

Therefore, the present invention relates to a glyceride mixture in whichthe content of conjugated fatty acids is higher than the content ofconjugated fatty acids of the starting glyceride mixture, and thefurther composition, for example of the glycerides, saturated fattyacids and/or esters of fatty acids, essentially corresponds to thecomposition of the starting glyceride mixture.

The term “essentially correspond to the composition of the startingglyceride” also includes changes as occur when the inventive process iscarried out, for example on account of the increased pressure, thereaction with a solvent or the high temperature, without addition ofcatalyst. The term “starting glyceride mixture” is also taken to mean,in particular, natural and complex synthetic oils and fats, preferablyrich in glycerides, in particular in triglycerides. However, themixtures can also have a content of free fatty acids and otheradditives, as are listed above, for example. To prepare CLA, the“starting glycerides” are rich in linoleic acid. Examples of preferrednatural oils are enumerated above. The term “starting glyceride mixture”also includes mixtures of different oils and fats.

Of economic advantage for the isolation of CLA is a glyceride mixture asproduct in which the content of conjugated linoleic acid is at least30%, preferably 50%, more preferably 60%, 70% or 80%, most preferablymore than 90%, of the content of linoleic acid of the starting glyceridemixture. However, small amounts of CLA in the oils or fats can also beadvantageous, for example in food additives.

Preferably, the inventive glyceride mixture has contents of11,13-octadecadienoic acid isomers, 8,10-octadecadienoic acid isomers,cis,cis-octadecadienoic acid isomers and/or trans/trans-octadecadienoicacid isomers in each case less than 5%, more preferably less than 3%,most preferably 1% or less, of the fatty acid content.

Particular preference is given to glyceride mixtures in which the CLAcontent in the triglyceride is greater than 30%, more preferably greaterthan 50%, 60% or 70%, most preferably greater than 80% or 90%, and whichhave a content of less than 5%, more preferably less than 3%, mostpreferably 1% or less, of 11,13-octadecadienoic ester isomers,8,10-octadecadienoic ester isomers, cis,cis-octadecadienoic esterisomers and/or trans/trans-octadecadienoic ester isomers. The CLAcontent in the triglyceride of greater than 50% consists hereprincipally of C18:2 cis-9, trans-11 and C18:2 trans-10, cis-11 CLA in aratio of 1:1.

Further preference is given to a glyceride mixture in which the CLAcontent in the triglyceride essentially consists of cis-9, trans-11 andtrans-10, cis-12 CLA.

The isoforms cis-9, trans-11 and trans-10, cis-12 CLA appear to be thebiologically active isoforms of CLA, therefore mixtures which have ahigh content of these two isoforms and a lower content of those isoformswhich have been enumerated above are particularly advantageous. Theinventive mixtures are particularly suitable for those applications inwhich high contents of CLA are advantageous, for example to producecosmetics, drug, food or animal feed preparations or a remedy.

In a further embodiment the present invention relates to a cosmetics,food, food supplement, animal feed or drug preparation which comprisesan inventive product, in particular the inventive alkene, for example asglyceride or glyceride mixture. Inventive alkenes, for exampleglycerides, can only be added such that the content of catalyst isharmless for the intended usage.

Said preparation can therefore contain additives.

The term “additives” is taken to mean further additives which areadvantageous for nutrition or health, for example “nutrients” or “activecompounds”. The preparation can comprise one or more additives foranimal or human nutrition or treatment and can be diluted or mixedtherewith. Additives can be administered together with, or separatelyfrom, the feed, food, food supplement or drug. A food, food supplement,animal feed or drug preparation contains no additives, or no quantitiesof additives, which may be considered harmful for animal or humannutrition.

“Nutrients” are those additives which are advantageous for the nutritionof humans or animals. Preferably, the inventive preparation thereforealso comprises vitamins, for example vitamins A, B₁, B₂, B₆, B₁₂, C, D₃,K and/or E, pantothenic acid, biotin, choline, folic acid, nicotinicacid, taurine, carboxylic acids, for example tricarboxylic acids,citrate, isocitrate, trans/cis aconitate, and/or homocitrate, enzymes,for example phytases, carotenoids, minerals, for example P, Ca, Mgand/or Na, proteins, carbohydrates, fats, amino acids and/or traceelements Mn, Zn, Cu, Co, Se, Fe and/or Cr. The preparation can alsocomprise pyruvic acid, L-carnitine, carbohydrases, lipoic acid,coenzymes Q10, aminocarboxylic acids, for example creatine.

“Active compounds” are substances which support the use of the inventivepreparation as drug or serve for their action in the treatment ofdisorders, in particular in the treatment of cancer, diabetes,overweight, AIDS, allergies and cardiovascular disorders (see alsobelow).

Therefore the inventive preparation can also include preservatives,antibiotics, antimicrobial additives, antioxidants, chelating agents,inert gases, physiologically harmless salts etc. Those skilled in theart know the additives suitable to add to the preparation for therespective use as drug, animal feed, food supplement or food additive,or can determine them by simple tests known in the prior art.

“Additives” are also antioxidants. Antioxidants are advantageous, forexample, to protect the double bonds of the fatty acids from oxidation.However, the general health-promoting action of antioxidants is alsoknown. Thus, in animal nutrition, ethoxyquin, BHT and/or BHA are used asantioxidants, otherwise, gamma- and alpha-tocopherols, tocotrienol,rosemary extract, naturally occurring polyphenols, for exampleflavonoids, isoflavones and carotenoids, are also used.

In a further embodiment, said preparation comprises furtherpolyunsaturated fatty acids (PUFAs). Preferably, the preparationcomprises omega-3-fatty acids, for example alpha-linolenic acid,docosahexaenoic acid, docosapentaenoic acid, and/or eicosapentaenoicacid, dimorphecolic acid, parinaric acid, and/or calendic acid, and/oromega-6-fatty acids, for example linoleic acid, gamma-linolenic acid,and/or dihomo-gamma-linolenic acid.

The inventive preparation can be solid, for example readily soluble inwater or oils, or liquid. The preparation, depending on use, has theappropriate dosage form, for example for animal nutrition, as foodadditive or as drug. Such dosage forms are, for example, tablets,capsules, powders, granules, sugar-coated tablets, solutions,nutrient-defined/balanced diets, such as enteral formula, andpreparations for infant nutrition, fat emulsions for parenteralnutrition, etc. Advantageous dosage forms of the preparations for therespective application are known to those skilled in the art. The fattyacids can be in free form, present as ethyl or methyl esters orpreferably, however, as triglyceride.

Flavorings can also be added to said preparations.

In foods, the preparation can be combined with customary foodcomponents. These include vegetable and animal products, in particularsugars, if appropriate in the form of syrups, fruit preparations, suchas fruit juices, nectar, fruit pulps, purees or dried fruits; cerealproducts and starches of said cereals; milk products, such as milkprotein, whey, yoghurt, lecithin and lactose.

The inventive preparation is also suitable for use in animal nutritionand can comprise, for example, additives.

“Additives” are taken to mean substances which serve for improvingproduct properties, such as dust behavior, flow properties, waterabsorption capacity and storage stability. Examples of such additivesand/or mixtures thereof can be those based on sugars, for examplelactose or maltodextrin, based on cereal products or legume products,for example corncob grit, wheat bran and soybean meal, based on mineralsalts, inter alia calcium salts, magnesium salts, sodium salts,potassium salts, and also D-pantothenic acid or its salts themselves(salt of D-pantothenic acid prepared chemically or by fermentation).

The present invention further relates to preparations which compriseinactive, viable and/or growing contents of organisms producingtriglycerides or other additives. Preferably, these are microorganisms,preferably fungi, yeasts and/or bacteria. Particularly preferably, theinventive-animal-feed comprises inactive, viable and/or growing contentsof fungi of the genus Mucor, yeasts of the genus Saccharomyces and/orbacteria of the Enterobacteriaceae, such as E. coli, Proteus vulgaris,Pseudomonads, such as Pseudomonas matophila, Bacillaceae, such asBacillus subtilis or Bacillus cereus, Coryneform bacteria, such asCorynebacterium glutamicum or Brevibacterium breve and/or Actinummycetalis and/or mixtures thereof. Very particular preference is givento bacteria of the genus Bacillus and, in this case, the speciesBacillus subtilis. Also, genetically modified and/or transgenicorganisms and/or production strains suitable for producing thepolyunsaturated fatty acids are included in the invention. Theenumeration above is not limiting here for the present invention.

If the inventive products, in particular the inventive alkene orglyceride mixtures are administered individually or in combination infeeds, the active compounds are administered as pure substance or asmixtures of substances or liquid or solid extracts together withcustomary feed constituents. Examples of customary feed constituentsare: corn, barley, wheat, oats, rye, triticale, sorghum, rice and brans,semolina brans and flours of these cereals, soybeans, soybean productssuch as soybean extraction meal, rapeseed, rapeseed extraction meal,cottonseed and extraction meal, sunflower seed, sunflower seedextraction meal, linseed, linseed extraction meal, expeller meals of oilseeds, field beans, peas, gluten, gelatin, tapioca, yeasts, single cellprotein, fish meal, salts, minerals, trace elements, vitamins, aminoacids, oils/fats and the like. Advantageous compositions are described,for example, in Jeroch, H. et al. Ernährung landwirtschaftlicherNutztiere [Farm animal nutrition], Ulmer, UTB.

The inventive preparation can be powder, granules, pellet, coatedextrudate and/or as a combination thereof. The preparation of theinventive animal feed, for example by means of coating compounds,serves, for example, for improving product properties, such as dustbehavior, flow properties, water absorption capacity and storagestability. Such preparations are extensively known in the prior art.Thus, in animal nutrition, for example, blocks of a solid, cohesiveshape-retaining mix of several kilos are used.

Animal nutrition preparations are composed in such a manner as to coveroptimally the appropriate nutrient requirement for the respective animalspecies. Generally, plant feed components such as cornmeal, wheat mealor barley meal, soybean whole meal, soybean extraction meal, linseedextraction meal, rapeseed extraction meal, green flour or ground peas,are chosen as sources of crude protein. In order to ensure anappropriate energy content of the feed, soybean oil or other animal orvegetable fats are added. Since the plant protein sources contain someessential amino acids only in an inadequate amount, feeds are frequentlyenriched with amino acids. This is primarily with lysine and methionine.In order to ensure the mineral and vitamin supply of the farm animals,in addition, minerals and vitamins are added. The type and amount ofadded minerals and vitamins depends on the animal species and is knownto those skilled in the art (see, for example, Jeroch et al., Ernährunglandwirtschaftlicher Nutztiere, Ulmer, UTB). To cover the nutrient andenergy requirements, complete feeds can be used which comprise allnutrients in a ratio to one another covering requirements. It can formthe sole feed of the animals. Alternatively, a supplementary feed can beadded to a cereal grain feed. This relates to protein-, mineral- andvitamin-rich feed mixtures that usefully supplement the grain feed.

In addition, the invention relates to an inventive preparation that is adrug. The combinations mentioned herein can advantageously be used inthe manufacture of drugs for treating cancer, allergies, diabetes and/orcardiovascular disorders, for example arteriosclerosis. A process forproducing cosmetics, drugs or remedies can comprise one of the steps ofthe inventive process and formulation of the process product into apharmacologically or dermatologically compatible form. Also, aninventive alkene, glyceride or glyceride mixture can be formulated intoa pharmacologically or dermatologically compatible form.

Improved food conversion, as has been observed for CLA, can lead to morerapid convalescence, for example, in the case of persons or animalsweakened by illness. The drugs prepared using the inventive preparationcan therefore also be used for the treatment of cancer, cardiovasculardisorders, for example arteriosclerosis (MacDonald, J. J. AmericanCollege of Nutrition, (2000) 19, 111S-118S), diabetes (WO99/29317),allergies, overweight and for disorders accompanying diets.

Thus, for example, the use of said preparation to accelerate build-up ofthe body, for example after a relatively long illness, which isaccompanied by loss of weight, for example chemotherapy, and to supportor accelerate the convalescence process is advantageous.

The drug can, in addition, comprise other active compounds, for examplethe abovementioned or other active compounds. The active compounds canbe used to treat cancer, cardiovascular disorders, for examplearteriosclerosis, diabetes, allergies, and to support diets or improvethe action of the inventive preparation. A drug for treating diabetescan comprise, for example, insulin, sulfonylureas, sulfonamides, lipoicacid, gamma-glucosidase inhibitors, thiazolidinediones, metformin and/oracetylsalicylic acid. Cancers are treated, for example, by addingcytostatics, such as vinca alkaloids, alkylating agents, for examplechlorambucil, melphalan, thio-TEPA, cyclophosphamide, etc., by folicacid analogs, such as aminopterin or methotrexate, or by the addition ofimmunosuppressives, for example cyclophophosphamide and azathioprine,glucocorticoids, such as prednisolone, or cyclosporin. HIV infections orAIDS can be treated, for example, by administering reverse transcriptaseinhibitors and/or protease inhibitors. Allergies are treated, forexample, by stabilizing the mast cells, for example using chromoglyxate,by blockading the histamine receptors, for example by H1-antihistamines,or by functional antagonists of the allergy mediators, for example byalpha-sympathomimetics, adrenalin, beta2-sympathomimetics, theophylline,ipratropium or glucocorticoids. Cardiovascular disorders are treatedusing coagulation inhibitors, ACE inhibitors, cholesterol-loweringagents, such as statins and fibrates, niacin, cholestyramine.

The drug can comprise a pharmaceutically compatible carrier. Examples ofsuitable pharmaceutically compatible carriers are known in the prior artand include physiologically harmless salts, for examplephosphate-buffered salines, water, emulsions, for example oil/wateremulsions, sterile solutions, etc. Sterile solutions can be, forexample, aqueous or non-aqueous solutions. Aqueous solutions are, forexample, water, alcohol/water solutions, emulsions or suspensions, andinclude sodium chloride solutions, Ringers dextrose, dextrose and sodiumchloride etc. Examples of non-aqueous solutions are propylenes, glycol,polyethylene glycol, vegetable oils, organic esters, for example ethyloleate. In addition, the drug can comprise one of the abovementionedsuitable additives.

Drugs can be administered in a conventional manner orally orparenterally (subcutaneously, intravenously, intramuscularly,intraperitoneally). They can also be administered via the nasal/throatcavity via vapors or sprays.

The dosage depends on age, condition and weight of the patient, and alsoon the type of administration. Generally, the daily active compounddosage is from about 0.05 to 100 mg/kg of body weight for oraladministration and from about 0.01 to 20 mg/kg of body weight forparenteral administration. Particular preference is given to from 0.5 to50 mg/kg.

The novel compounds can be used in the solid or liquid state in theconventional pharmaceutical dosage forms, for example as tablets, filmtablets, capsules, powder, granules, sugar-coated tablets,suppositories, solutions, lotions, creams or sprays. These aremanufactured in a conventional manner. The active compounds in this casecan be processed together with the customary pharmaceutical aids, suchas tablet binders, fillers, preservatives, tablet disintegrators,viscosity controlling agents, emollients, wetting agents, dispersants,emulsifiers, solvents, retarding agents, antioxidants and/or propellantgases (see H. Sucker et al.: Pharmazeutische Technologie [Pharmaceuticaltechnology], Thieme-Verlag, Stuttgart, 1991). The resultant dosage formscomprise active compounds usually in an amount from 0.1 to 90% byweight.

An inventive drug can be manufactured, for example, by producing crudeextracts from plants that comprise unsaturated esterified fatty acids,in particular triglycerides, for example in the form of theabovementioned oils and fats, reacting them and formulating them.Standard manufacturing processes for drugs are sufficiently known tothose skilled in the art.

Depending on purpose, the amount of glyceride used, for example the CLAused, must be adapted. The amount of glyceride used can be, for example,0.01 or 0.1% of the amount of fat added during feeding. Preference isalso given to 0.5%, 1%, 2% or 3%, 5% or 10% glyceride, in particularCLA.

In the case of animal nutrition preparations, the proportion of theoils/fats added to the amount of fat during feeding can be up to 100%.If the amount of fat added is based on the fat content analyzed in thefeed (total fat), the proportion of CLA can be 75% or less.

In a further embodiment the present invention also relates to a kitwhich comprises the inventive preparation. The preparation can be packedin one or more containers. The constituents of the inventivepreparation, for example the alkene, glyceride or glyceride mixture, butalso the abovementioned additives, can be packed separately or togetherin one container of the kit. The kit can be used for carrying out theinventive process and contain instructions for carrying it out.

Various documents are cited in the present text of this description.Each of the documents (including instructions and descriptions ofmanufacturers) is hereby incorporated in the description by reference.This does not mean, however, that each of said documents is actuallyprior art for the present invention.

The present invention is explained by the following examples anddrawings, without these restricting this present invention in anymanner.

EXAMPLES

-   1. 100 g of thistle oil (Cereol, contains 72% of linoleic acid) are    heated with 10 g of 1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD) under    argon for 6 hours at 135° C. The mixture is allowed to come to room    temperature and the solution is made acid using 1N HCl. The reaction    solution is washed twice with 350 ml of water. The remaining organic    phase is taken up in dichloromethane and concentrated at 50° C. and    7 mbar on a rotary evaporator. Since the resultant oil, in the form    of the triglyceride, cannot be analyzed by GC, it is reacted with    potassium ethoxide in ethanol to give the ethyl esters of the fatty    acids.    -   GC result (fatty acids as ethyl esters): palmitic acid 6.8%;        stearic acid 3.1%, oleic acid 14.5%, 9-cis,11-trans conjugated        linoleic acid 35.4%, 10-trans,12-cis conjugated linoleic acid        35.8%.-   2. In a reaction block, in each case 10 g of the ethyl ester of a    fatty acid, produced by transesterification of sunflower seed oil,    having a content of 9cis,12cis-linoleic acid ethyl ester of 66% were    heated with 1 g of the respective base at 130° C. The product was    analyzed by GC. The percentage peak areas are shown.

LA EE [area CLA EE Catalyst %] [area %]2-tert-Butyl-1,1,3,3-tetramethylguanidine 65.7 0.0 (Barton base)1,5,7-Triazabicyclo(4.4.0)dec-5-ene 0.0 63.51,3,4,6,7,8-Hexahydro-1-methyl-2H- 65.9 0.0 pyrimidino-(1,2A)-pyrimidine2-tert-Butyl-2-diethylamino-1,3- 65.9 0.0dimethylperhydro-1,3,2-diazaphosphorine

-   3. 9cis,12cis-Linoleic acid ethyl ester (Aldrich, 98%), and    trilinolein (Aldrich) were reacted with various bases: analysis was    performed by GC. The peak areas are shown.

Proportion of base to CLA CLA Starting fatty acid Reaction AnalysisStarting Linoleic CLA CLA cis/ trans/ No. material Base residue Solventtime/temp. as material acid 9c11t 10t12c cis trans 1 LA-EE Phosphazenebase 120 mol %  THF  30 min at 30° C. Ethyl ester 28.1 — 32.7 36.1 0.60.6 P4-t-Bu solution (1M in hexane)  60 min at 30° C. Ethyl ester 11.1 —40.3 44.9 0.7 0.8  30 min at 0° C. Ethyl ester — — 45.1 51.5 0.8 1.5  60min at 0° C. Ethyl ester — — 45   51   0.8 2.1  30 min at 20° C. Ethylester — — 43.7 49.4 0.8 5.1 120 min at 20° C. Ethyl ester — — 37.9 43.91   16 2 LA-EE Phosphazene base P4-Bu 10 mol % without 1 h at 0° C.Ethyl ester — — 46   48.8 0.9 1.4 solution (1M in hexane) solvent 2 h at0° C. Ethyl ester — — 46.4 49.8 1   2 1 h at 20° C. Ethyl ester — — 42.345.9 1   3.6 2 h at 20° C. Ethyl ester — — 45.2 48.8 1   4.1 1 h at 80°C. Ethyl ester — — 44.9 48.5 1   4.9 1 h at 120° C. Ethyl ester — — 44.848.4 1   4.9 3 LA-EE 1,8-Diazabicyclo[5.4.0]- 10 mol % without 1 h at120° C. Ethyl ester 98.6 — — — — — undec-7-ene solvent 4 TrilinoleinPhosphazene base P4-Bu 10 mol % without 1 h at 0° C. Ethyl ester 73.5 —11.4 13.7 — — solution (1M in hexane) solvent 5 LA-EE 1,8-Bis(dimethyl-10 mol % without 1 h at 0° C. Ethyl ester 99.4 — — — — —amino)naphthalene solvent 1 h at 20° C. Ethyl ester 99.2 — — — — — 2 hat 20° C. Ethyl ester 99.3 — — — — — 1 h at 80° C. Ethyl ester 99.5 — —— — — 1 h at 120° C. Ethyl ester 99 — — — — — 6 LA-EE1,3,4,6,7,8-Hexahydro- 10 mol % without 1 h at 0° C. Ethyl ester 99.3 —— — — — 1-methyl-2H-pyrimido- solvent [1,2-a]pyrimidine 1 h at 20° C.Ethyl ester 99.2 — — — — — 2 h at 20° C. Ethyl ester 99.3 — — — — — 1 hat 80° C. Ethyl ester 99.6 — — — — — 1 h at 120° C. Ethyl ester 99 — — —— — 7 LA-EE 1,5,7-Triazabicyclo- 10 mol % without 1 h at 0° C. Ethylester 99.5 — — — — — [4.4.0]dec-5-ene solvent 8 Trilinolein Phosphazenebase P4-Bu 10 mol % without 1 h at 20° C. Ethyl ester 39.7 — 19.8 24   —— solution (1M in hexane) solvent 9 Trilinolein 1,5,7-Triazabicyclo- 10mol % toluene 4 h at 110° C. Ethyl ester 71.6 — 10.4 12.5 — —[4.4.0]dec-5-ene 10 Trilinolein 1,5,7-Triazabicyclo- 10 mol % toluene 4h at 110° C. Ethyl ester 96.2 — — — — — [4.4.0]dec-5-ene bound topolystyrene 11 LA-EE 1,8-Diazabicyclo- 10 mol % toluene 2 h at 110° C.Ethyl ester 99.2 — — — — — [5.4.0]undec-7-ene 4 h at 110° C. Ethyl ester99.5 — — — — — 6 h at 110° C. Ethyl ester 99.4 — — — — — 12 LA-EE1,5,7-Triazabicyclo- 10 mol % toluene 2 h at reflux Ethyl ester 76.4 —10.7 12   — — [4.4.0]dec-5-ene 4 h at reflux Ethyl ester 60 — 18.4 20.1— — 6 h at reflux Ethyl ester 47.6 — 24.3 26.1 — — 13 LA-EE1,3,4,6,7,8-Hexahydro- 10 mol % toluene 2 h at reflux Ethyl ester 99.4 —— — — — 1-methyl-2H-pyrimido- [1,2-a]pyrimidine 4 h at reflux Ethylester 97.8 — — — — — 6 h at reflux Ethyl ester 98.9 — — — — —

-   4. Linoleic acid methyl ester (66%) is continuously reacted with    1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD) in a tube at 150° C. to    give conjugated linoleic acid methyl ester:    -   440.7 g of linoleic acid methyl ester (65.8%) and 49.9 g of TBD        (Fluka, 98%) are charged into a flask under argon at room        temperature and pumped through a steel coil (internal diameter        8 mm) heated to 150° C. The dead volume in the steel coil is 90        g.

Linoleic CLA-M CLA-M Cis, cis-CLA Trans, trans-CLA acid ethyl E 9c11t E10t12c isomers isomers Reaction time in No. ester [area %] [area %][area %] [area %] the steel coil 1 31.5 16.9 16.9 0.7 0.3 approximately26 min. 2 17.8 23.7 23.6 1.0 0.4 approximately 68 min.

-   5. Repetition of the transesterification of vegetable oil with    1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD) according to the    instructions of Schuchardt et al., J. Braz. Chem. Soc., Vol. 9, No.    3, 199-210, 1998:    -   80.1 g (0.091 mol, equivalent to 0.27 mol of fatty acids) of        sunflower seed oil (Bonita) are heated at 70° C. for 2 hours        with 20 g (0.62 mol) of methanol and 0.31 g (0.002 mol,        equivalent to 1 mol %) of 1,5,7-triazabicyclo[4.4.0]dec-5-ene        (Fluka). The transesterification was followed using GC. No        conjugated linoleic acid (CLA) was formed.    -   Analysis:

Reaction time Palmitic acid Stearic acid Oleic acid Linoleic acid (min)C16:0 C18:0 C18:1 C18:2 CLA 20 6.1 4.6 23.0 63.8 n.d. 60 6.1 5.1 22.863.9 n.d. 90 6.1 4.4 23.5 64.0 n.d. 120  6.1 4.8 23.1 64.0 n.d. n.d. notdetectable

-   6. Linoleic acid methyl ester (64.4%) is continuously reacted with    1,5,7-triazabicyclo[4.4.0.]-dec-5-ene (TBD) in a tube at 150° C. to    give conjugated linoleic acid methyl ester:    -   223.8 g of linoleic acid methyl ester (64.4%) and 25.3 g of TBD        (Fluka, 98%) are charged into a flask under argon at room        temperature and pumped through a steel coil (internal diameter        2 mm) heated to 150° C. The dead volume in the steel coil is        11.7 mL. This corresponds to approximately 9.4 g. The pumping        rate was set at 3 g/h.

CLA- CLA- Linoleic ME ME Cis, cis- Reaction acid 9c11t 10t12c CLA cis,cis-CLA Product time in the Time Time methyl [Area [Area isomers isomersweight steel coil No. from to ester %] %] [Area %] [Area %] [g] [h] 1 7:40  8:30 20.8 20.7 0.9 0.3 — — 2  8:30  9.30 26.4 26.3 1.0 0.3 6.5 —3  9:30 10:30 29.0 28.9 1.2 0.4 3.3 3 4 10:30 11:30 30.1 30.0 1.3 0.02.1 4 5 11:30 12:30 29.9 29.8 1.2 0.4 3.0 3

-   7. Linoleic acid methyl ester (64.4%) is heated with    1,5,7-triazabicyclo[4.4.0.]-dec-5-ene (TBD) and toluene under reflux    (115° C.).    -   11.2 g of linoleic acid methyl ester (64.4%; 0.025 mol), 1.05 g        of TBD (Fluka, 98%; 0008 mol) and 11.2 g of toluene (0.12 mol)        are charged into a flask under argon at room temperature and        then heated under reflux (115° C.).

Linoleic Reaction acid CLA-ME CLA-ME Cis, cis-CLA Trans, trans- timemethyl 9c11t 10t12c isomers CLA isomers [h] ester [Area %] [Area %][Area %] [Area %] 1 60.8 2.7 2.6 — — 2 56.3 4.4 4.4 — — 3 49.1 8.5 8.4 —— 4 43.3 12.0 11.8 — — 5 36.9 14.9 14.8 — — 6 31.5 17.5 17.4 — —

-   8. Linoleic acid methyl ester (64.4%) is heated with    1,5,7-triazabicyclo[4.4.0.]-dec-5-ene (TBD) and xylene (mixture of    o, m and p-xylene) under reflux (145° C.).    -   11.5 g of linoleic acid methyl ester (64.4%; 0.026 mol), 1.04 g        of TBD (Fluka, 98%; 0.008 mol) and 11.3 g of xylene (0.11 mol)        are charged into a flask under argon at room temperature and        then heated under reflux (145° C.).

Linoleic Reaction acid CLA-ME CLA-ME Cis, cis-CLA Trans, trans- timemethyl 9c11t 10t12c isomers CLA isomers [h] ester [Area %] [Area %][Area %] [Area %] 1 34.9 16.0 15.9 — — 2 16.3 24.7 24.6 — — 3 9.4 28.128.0 — — 4 5.0 30.4 30.2 — — 5 2.6 31.5 31.3 — — 6 1.5 32.0 31.8 — —

-   9. Linoleic acid methyl ester (64.4%) is heated to 140° C. with    1,5,7-triazabicyclo[4.4.0.]-dec-5-ene (TBD) and diethylene glycol    dimethyl ether.    -   11.6 g of linoleic acid methyl ester (64.4%; 0.026 mol), 1.05 g        of TBD (Fluka, 98%; 0.008 mol) and 11.5 g of DEGDM (0.09 Mol)        are charged into a flask under argon at room temperature and        then heated to 140° C.

Linoleic acid CLA-ME CLA-ME Cis, cis-CLA Trans, trans- Reaction methyl9c11t 10t12c isomers CLA isomers time [h] ester [Area %] [Area %] [Area%] [Area %] 1 40.9 9.7 9.6 — — 2 19.5 21.1 21.0 — — 3 10.9 25.4 25.3 — —4 6.8 26.0 25.9 — — 5 5.6 28.6 28.5 — — 6 4.2 28.8 28.7 — —

-   10. Linoleic acid methyl ester (64.4%) is heated with    1,5,7-triazabicyclo[4.4.0.]-dec-5-ene (TBD) and mesitylene under    reflux (168° C.).    -   11.6 g of linoleic acid methyl ester(64.4%; 0.026 mol), 1.03 g        of TBD (Fluka, 98%; 0.007 mol) and 11.8 g of DEGDM (0.1 mol) are        charged into a flask under argon at room temperature and then        heated to 168° C.

Linoleic Cis, cis- and Reaction acid CLA-ME CLA-ME trans, trans- timemethyl 9c11t 10t12c CLA isomers [h] ester [Area %] [Area %] [Area %] 127.2 18.8 18.8 0.4 (only t, t) 2 3.5 30.0 29.9 1.3 3 0.8 31.1 31.0 2.2 40.7 30.9 30.8 2.6 5 0.5 30.4 30.4 2.6 6 0 27.8 27.6 too dilute

1. A process for preparing a C₈-C₂₄ alkene or alkene derivative having aconjugated double bond, which comprises isomerizing an alkene having twodouble bonds separated by a methylene bridge with an a) imino base, orb) iminophosphorane base, as catalyst.
 2. A process as claimed in claim1, wherein a) is an aminoimino base and b) is an aminoiminophosphoranebase.
 3. A process as claimed in claim 1, wherein the alkene having twoseparated double bonds is a polyunsaturated fatty acid ester (FAE II),and the alkene having a conjugated double bond is a conjugatedunsaturated fatty acid ester (FAE I).
 4. A process as claimed in claim3, wherein the fatty acid ester (FAE II) is an alkyl ester.
 5. A processas claimed in claim 3, wherein the fatty acid ester (FAE II) is aglyceride.
 6. A process as claimed in claim 5, wherein the glyceride ispresent in a synthetic or natural glyceride mixture.
 7. A process asclaimed in claim 5, wherein the glyceride mixture is of animal,microbial or vegetable origin, or a derivative or a mixture thereof. 8.A process as claimed in claim 5, wherein the glyceride is atriglyceride.
 9. A process as claimed in claim 1, wherein linoleic esteris converted to conjugated linoleic ester.
 10. A process as claimed inclaim 1, wherein C18:2 cis-9, trans-11 and/or C18:2 trans-10, cis-12CLAs are produced.
 11. A process as claimed in claim 1, wherein thecatalysis is carried out using compound (I)

where independently of one another X₁ can be —NH— or —PH—, X₂ can be C,N or P; and where R1 to R4 independently of one another can be: H,branched or unbranched C₁- to C₂₀-alkyl, where from 0 to 3 carbon atomscan be replaced by O, S, NZ and/or —X₃—(C═X₄)—, mono-, bi-, ortricyclic, aromatic, saturated or partially unsaturated C₀- toC₆-alkylcarbocycle or heterocycle having from 3 to 17 carbon atoms,where from 0 to 3 heteroatoms can be selected from O, S, NZ and/or—X₃—(C═X₄)—; and where each carbon atom of the alkyl chains or of thering can bear up to three of the following substituents OZ, SZ,(C═O)—OZ, NZZ₁, C₁- to C₆-alkyl; and where X₃ can be a bond, O, S or NZ,and/or X₄ can be O, S or NZ; and where Z and/or Z₁ independently of oneanother can be H or C₁- to C₆-alkyl; and where R1 and R4 and/or R2 andR3 can be part of a ring.
 12. A process as claimed in claim 1, whereinthe catalysis is carried out using compound (II)

wherein R and R1 to R6 independently of one another can be: H, branchedor unbranched C₁- to C₂₀-alkyl, where from 0 to 3 carbon atoms can bereplaced by O, S, NZ and/or —X₃—(C═X₄)—; mono-, bi-, or tricyclic,aromatic, saturated or partially unsaturated C₀- to C₆-alkylcarbocycleor heterocycle having from 3 to 17 carbon atoms, where from 0 to 3heteroatoms can be selected from O, S, NZ and/or —X₃—(C═X₄)—; and whereeach carbon atom of the alkyl chains or of the ring can bear up to threeof the following substituents OZ, SZ, (C═O)—OZ, NZZ₁, C₁- to C₆-alkyl;where X₃ can be a bond, O, S or NZ, and/or X₄ can be O, S or NZ; where Zand/or Z₁ independently of one another can be H or C₁- to C₆-alkyl; andwhere different R1 to R6 can also be part of one or of differentring(s).
 13. A process as claimed in claim 1, wherein the catalysis iscarried out using 1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD),1,2,3,4,4a,5,6,7-octahydro-1,8-naph-thyridine [CAS 60832-40-8],phosphazene base P4-T-BU [CAS 111324-04-0], phosphazene base P1-T-OctNo. [CAS 161118-69-0], phosphazene base P1-T-Bu-tris(tetramethylene)[CAS 161118-67-8], phosphazene base P2-T-Bu [CAS 111324-03-9],phosphazene base P4-T-Oct [CAS 153136-05-1], their salts, or the salts1,1,1,3,3,3-hexakis (dimethylamino)diphosphazenium fluoride [CAS137334-99-7], 1,1,1,3,3,3-hexakis(dimethylamino)diphosphazeniumtetrafluoroborate [CAS 137334-98-6]or2-tert-butylimino-2-die-thylamino-1,3-dimethylperhydro-1,3,2-diazaphosphorine[CAS 98015-45-3], or their bases.
 14. A process as claimed in claim 1,wherein the catalyst is separated off.
 15. A composition comprising aconjugated alkene, a fatty acid, a fatty acid ester, a glyceride, atriglyceride or mixtures thereof obtained by the process defined inclaim 1, and a part or all of the catalyst employed in said process. 16.A glyceride mixture to obtained by subjecting a glyceride mixture ofanimal, microbial or vegetable origin and comprising at least oneglyceride having two double bonds separated by a methylene bridge to theprocess claim
 1. 17. A glyceride mixture wherein the content ofconjugated fatty acids in the glyceride mixture is higher than thecontent of conjugated fatty acids of the starting glyceride mixture andthe composition of the further glycerides essentially corresponds to thecomposition of a glyceride mixture as defined in claim
 7. 18. An alkene,a fatty acid, a fatty acid ester, a glyceride, a triglyceride or amixture thereof as claimed in claim 15, wherein the content ofconjugated linoleic esters is at least 30% of the content of linoleicesters of the starting glyceride mixture.
 19. An alkene, a fatty acid, afatty acid ester, a glyceride, a triglyceride or a mixture thereof asclaimed in claim 15, wherein the content of 11,13-octadecadienoic acidisomers, 8,10-octadecadienoic acid isomers, cis,cis-octadecadienoic acidisomers or trans/trans-octadecadienoic acid isomers is in each case lessthan 5% of the fatty acid content.
 20. An alkene, a fatty acid, a fattyacid ester, a glyceride, a triglyceride or a mixture thereof as claimedin claim 15, wherein the CLA content in the triglyceride essentiallyconsists of cis-9, trans-11 and trans-10, cis-12 CLA.
 21. A foodpreparation, a food supplement preparation, an animal feed preparation,a drug preparation or a cosmetics preparation comprising the alkene, thefatty acid, the fatty acid ester, the glycerides, the triglycerides ormixtures thereof as claimed in claim
 15. 22. A drug for treatingallergies, diabetes, cancer, cardiovascular disorders or obesitycomprising an alkene, a fatty acid, a fatty acid ester, a glyceride or atriglyceride as claimed in claim
 15. 23. A process as claimed in claim7, wherein the glyceride mixture is olive oil, canola oil, coconut oil,coconut fat, sesame seed oil, rice germ oil, bamboo oil, bamboo fat,sunflower seed oil, rapeseed oil, fish oil, tallow oil, soybean oil,palm oil, safflower oil, linseed oil, wheatgerm oil, peanut oil,cottonseed oil, corn oil, pig fat, beef fat, poultry fat, milk fat, tungoil or shea oil, or a derivative or a mixture thereof.
 24. A glyceridemixture wherein the content of conjugated fatty acids in the glyceridemixture is higher than the content of conjugated fatty acids of thestarting glyceride mixture and the composition of the further glyceridesessentially corresponds to the composition of the glyceride mixturedefined in claim
 16. 25. The glyceride mixture defined in claim 16,wherein the content of conjugated linoleic esters is at least 30% of thecontent of linoleic esters of the starting glyceride mixture.
 26. Theglyceride mixture defined in claim 16, wherein the content of11,13-octadecadienoic acid isomers, 8,10-octadecadienoic acid isomers,cis,cis-octadecadienoic acid isomers or trans/trans-octadecadienoic acidisomers is in each case less than 5% of the fatty acid content.
 27. Theglyceride mixture defined in claim 16, wherein the CLA content in thetriglyceride essentially consists of cis-9, trans-11 and trans-10,cis-12 CLA.
 28. A food preparation, a food supplement preparation, ananimal feed preparation, a drug preparation or a cosmetics preparationcomprising the glyceride mixture defined in claim
 16. 29. A drug fortreating allergies, diabetes, cancer, cardiovascular disorders orobesity comprising the glyceride mixture defined in claim 16.